The possibility of genetically engineering poultry to make them resistant to avian influenza is attracting attention and has now become a real possibility with improved methods for genetic modification and the emergence of RNAi as an antiviral strategy. In order to test this possibility, we have generated transgenic mice that express RNAi molecules targeting a conserved region of the influenza A NP gene and are testing these mice for resistance to influenza infection. Transgenes were initially developed that express short hairpin RNAs (shRNAs) targeting multiple influenza A viral genes. The shRNAs were tested for inhibition of H1N1 PR8 virus in vitro. Two potent shRNAs that target the NP and PA genes were chosen for lentiviral mediated generation of transgenic mice. Transgenic founders for the NP shRNA construct and also a negative control shRNAtargeting EGFP were generated. The constitutive expression of the shRNA molecules in a range of tissue types including lung, was confirmed and so far stable transmission of the RNAi transgenes from the F0 to F3 generation has been observed. Resistance to influenza infection in these transgenic mice is now being confirmed.
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